LOCUS       sequence-455454-        7673 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid enabling yeast-mediated expression and secretion of Gaussia 
            luciferase (GLuc).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7673)
  AUTHORS   Rebeck ON, Wallace MJ, Prusa J, Ning J, Evbuomwan EM, Rengarajan S, 
            Habimana-Griffin L, Kwak S, Zahrah D, Tung J, Liao J, Mahmud B, 
            Fishbein SRS, Ramirez Tovar ES, Mehta R, Wang B, Gorelik MG, Helmink
            BA, Dantas G
  TITLE     A yeast-based oral therapeutic delivers immune checkpoint inhibitors
            to reduce intestinal tumor burden.
  JOURNAL   Cell Chem Biol. 2024 Nov 16:S2451-9456(24)00452-5. doi: 
            10.1016/j.chembiol.2024.10.013.
  PUBMED    39571582
REFERENCE   2  (bases 1 to 7673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Chem 
            Biol. 2024 Nov 16:S2451-9456(24)00452-5. doi: 
            10.1016/j.chembiol.2024.10.013."
COMMENT     Sequence Label: pOGS539_GLuc
FEATURES             Location/Qualifiers
     source          1..7673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      139..1481
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(1102..1149)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1741..2535
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1741..2535
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1795..1814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2405..2424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2548..2684
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2829..2900
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    2922..2970
                     /note="synthetic polyadenylation signal"
     promoter        3153..3566
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             join(3671..3730,3731..3928)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MGRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDL
                     EGDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKR"
     CDS             3941..3967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      4601..4848
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4617..4635)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      4906..4952
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4982..5118
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     promoter        5204..5723
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     misc_feature    6428..6499
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6725..7313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6805..6824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
       61 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
      121 tcgaaatcgg ggcgcgccaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
      181 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
      241 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
      301 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
      361 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
      421 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
      481 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
      541 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
      601 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
      661 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
      721 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
      781 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
      841 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
      901 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
      961 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     1021 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct
     1081 tttggttttc aaaagcgctc tgaagttcct atacttccta gagaatagga acttcggaat
     1141 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
     1201 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
     1261 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
     1321 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
     1381 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
     1441 gtctcatcct tcaatgctat catttccttt gatattggat catacggcgc gcctggtgta
     1501 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     1561 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     1621 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     1681 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     1741 atgattgaac aagatggatt gcacgcaggt tctccggcgg cttgggtgga gaggctattc
     1801 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1861 gcgcaggggc gtccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1921 caagacgagg cagcgcggct atcgtggctg gcgacgacgg gcgttccttg cgcggctgtg
     1981 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2041 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2101 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2161 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2221 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgtc tatgcccgac
     2281 ggcgaggatc tcgtcgtgac ccacggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2341 ggccgctttt ctggattcat cgactgtggc cgtctgggtg tggcggaccg ctatcaggac
     2401 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2461 cttgtgcttt acggtatcgc cgcgcccgat tcgcagcgca tcgccttcta tcgccttctt
     2521 gacgagttct tctgaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     2581 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     2641 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt ttaaactcga
     2701 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     2761 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     2821 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     2881 agcagcttca gctacctctc aatttcaatc gttgcgttac acacacaaaa aaccaacaca
     2941 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     3001 tcaatgcata ctttgtacgt tcaaaataca atgcagtaga tatatttatg catattacat
     3061 ataatacata tcacatagga agcaacaggc gcgttggact tttaattttc gaggaccgcg
     3121 aatccttaca tcacacccaa tcccccacaa gtgatccccc acacaccata gcttcaaaat
     3181 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     3241 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     3301 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     3361 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     3421 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     3481 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     3541 aagaaagcat agcaatctaa tctaagtttt aattacaaaa gatctttgtc gatcctacca
     3601 tccactcgac acacccgcca gcggccgctg ccaagcttcc gagctctcga attcaaagga
     3661 ggtaccaacc atggggagat ttccttcaat ttttactgca gttttattcg cagcatcctc
     3721 cgcattagct gctccagtca acactacaac agaagatgaa acggcacaaa ttccggctga
     3781 agctgtcatc ggttacttag atttagaagg ggatttcgat gttgctgttt tgccattttc
     3841 caacagcaca aataacgggt tattgtttat aaatactact attgccagca ttgctgctaa
     3901 agaagaaggg gtatctttgg ataaaagaga gggaagttcc tatccctatg acgtgccaga
     3961 ctacgcatcc agcggaggtg gatcaggtaa accgaccgag aacaatgaag atttcaacat
     4021 cgtcgccgta gcatcaaatt tcgccacgac ggacttagat gcagacagag gaaaactacc
     4081 tgggaaaaaa cttcccttgg aagtactgaa agaaatggag gcgaacgcga gaaaagctgg
     4141 ctgtaccagg ggttgtctta tatgtctgtc acatatcaaa tgtactccaa agatgaagaa
     4201 atttattccc gggcgttgcc atacatacga aggggataaa gagtcagctc aagggggaat
     4261 cggggaggcg atagttgata tacctgagat ccctggtttt aaggacctgg agcccatgga
     4321 gcaattcata gcgcaggtgg atttatgtgt ggactgcacc accggatgtc taaaaggcct
     4381 agctaacgta cagtgctccg acttactaaa aaaatggtta ccccagcgtt gtgcgacttt
     4441 tgcttcaaaa atccagggcc aggtggacaa aataaagggg gcgggaggcg attgatctag
     4501 aagttgtctc ctcctgcact gactgactga tacaatcgat ttctggatcc gcaggcctct
     4561 gctagcttga ctgactgaga tacagcgtac cttcagctca tcatgtaatt agttatgtca
     4621 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     4681 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     4741 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     4801 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccaaagcaag
     4861 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     4921 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     4981 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     5041 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     5101 ccagaaattt atccttaagg tcgtcagcta tcctgcaggg tgtttaaaga ttacggatat
     5161 ttaacttact tagaataatg ccattttttt gagttataat aatcctacgt tagtgtgagc
     5221 gggatttaaa ctgtgaggac cttaatacat tcagacactt ctgcggtatc accctactta
     5281 ttcccttcga gattatatct aggaacccat caggttggtg gaagattacc cgttctaaga
     5341 cttttcagct tcctctattg atgttacacc tggacacccc ttttctggca tccagttttt
     5401 aatcttcagt ggcatgtgag attctccgaa attaactaaa gcaatcacac aattctctcg
     5461 gataccacct cggttgaaac tgacaggtgg tttgttacgc atgctaatgc aaaggagcct
     5521 atataccttt ggctcggctg ctgtaacagg gaatataaag ggcagcataa tttaggagtt
     5581 tagtgaactt gcaacattta ctattttccc ttcttacgta aatatttttc tttttaattc
     5641 taaatcaatc tttttcaatt ttttgtttgt attcttttct tgcttaaatc tataactaca
     5701 aaaaacacat acataaacta aaaatggcca aaaccttcaa catctctcag caggatctgg
     5761 agctggtgga ggtcgccact gagaagatca ccatgctcta tgaggacaac aagcaccatg
     5821 tcggggcggc catcaggacc aagactgggg agatcatctc tgctgtccac attgaagcct
     5881 acattggcag ggtcactgtc tgtgctgaag ccattgccat tgggtctgct gtgagcaacg
     5941 ggcagaagga ctttgacacc attgtggctg tcaggcaccc ctactctgat gaggtggaca
     6001 gatccatcag ggtggtcagc ccctgtggca tgtgtagaga gctgatctct gactatgctc
     6061 ctgactgctt tgtgctcatt gagatgaatg gcaagctggt caaaaccacc attgaggaac
     6121 tcatccccct caagtacacc aggaactaag gttaatataa ttatataaaa atattatctt
     6181 cttttcttta tatctagtgt tatgtaaaat aaattgatga ctacggaaag cctttttata
     6241 ttgtttcttt ttcattctga gccacttaaa tttcgtgaat gttcttgtaa gggacggtag
     6301 atttacaagt gatacaacaa aaagcaaggc gctttttcta ataaaaagaa gaaaagcatt
     6361 taacaattga acacctctat atcaacgaag aatattcctg caggcgatct ctcgatttcg
     6421 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     6481 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     6541 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     6601 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     6661 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     7561 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     7621 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggc
//
